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Frozen autopsy brain tissue from fourteen multiple-sclerosis donors and three neurologically healthy controls was sectioned at –20 °C into 80–100 µm slices, and an 8 mm punch (~25 mg) of white matter or matched control tissue was collected from each region of interest . Nuclei were liberated with the 10x Genomics Chromium Nuclei Isolation Kit, using an extended lysis–debris-removal protocol tailored to lipid-rich white matter; the final suspension was stained with propidium iodide and counted ...

In this study, our experimental design focused on the detailed analysis of lesion areas in the brain tissues of patients with multiple sclerosis (MS). First, we collected brain tissue samples from 14 MS patients. Using specialized staining techniques, we accurately identified the locations of MS lesions within the brain tissues. Subsequently, we used a brain puncher to extract the brain tissue regions containing lesions from each sample. These precisely cut samples were then analyzed on the ...

8-week-old male Cx3cr1CreERT2/+; Abca1flox/flox; Abcg1flox/flox experimental animals (KO) and Cx3cr1CreERT2/+ controls (Ctrl) were used for EAE experiments. Myelin oligodendrocyte glycoprotein (MOG)-EAE was induced in mice purchased from Charles River Laboratories by subcutaneous immunization with 200 mg of MOG peptide 35–55 (MOG35–55) in complete Freund’s adjuvant (Mycobacterium tuberculosis at 3.75 mg ml−1; BD) and intraperitoneal injection twice with 500 ng of pertussis toxin (Sigma). Animals ...

Primary mouse microglia were purified from early postnatal brains, seeded on coated slides in defined medium, and exposed to a myelin plus IFN-γ activation regimen. After fixation and permeabilization, a 500-gene MERFISH panel was applied to spatially resolve lipid-metabolism and inflammatory transcripts at single-cell resolution, enabling unbiased identification of distinct microglial states across the culture.

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